Expand High Fidelity PCR System

for one-step RT-PCR amplification of up to 3 kb RNA targets

Cat. No. 1 732 641      100 units
Cat. No. 1 732 650      500 units
Cat. No. 1 759 078    2500 units

Description

Unique enzyme blend of Taq and Pwo DNA Polymerases. The Expand High Fidelity PCR System also includes:

Expand High Fidelity buffer, 10x conc., with 15 mM MgCl₂
Expand High Fidelity buffer, 10x conc., without MgCl₂
MgCl₂ stock solution, 25 mM

Application

Expand High Fidelity PCR System is an unique enzyme blend (Taq / Pwo DNA polymerase) that is ideal for amplifying fragments up to 5 kb from human genomic DNA. It gives better results (higher yield and greater fidelity) than Taq DNA Polymerase alone in standard PCR reactions. (See also "How the Expand High Fidelity PCR System Outperforms Taq DNA Polymerase").

Note: If Expand Reverse Transcriptase (Cat. No. 1785826)a, AMV Reverse Transcriptase (Cat. No. 1495062), or the First Strand cDNA Synthesis for RT-PCR (Cat. No. 1483188) is used for cDNA synthesis in a two step RT-PCR, RNA templates with a length up to 5 kb can be amplified. Expand Reverse Transcriptase is a genetically engineered version of M-MuLV Reverse Transcriptase. A point mutation within the RNase H sequence reduces the RNase H activity below detectable levels. This product is not for sale in the United States.

Application profile

Operating parameters

pH optimum: 9.2
Divalent ion requirement: Mg²⁺
Temperature optimum (for elongation): 68°–72°C

Experimental results

Figure 1: Expand High Fidelity successfully amplified the fragment from as little as 1 ng of genomic DNA.

Figure 2: Consistent amplification was achieved over the full range from 1.5 mM to 5 mM Mg2+ with Expand High Fidelity.

Key advantages

Improves fidelity of PCR amplification, because the enzyme blend amplifies with threefold greater accuracy than Taq DNA Polymerase alone
Produces more DNA with fewer cycles, because the proofreading activity of the enzyme blend reduces the number of truncated amplification products formed and increases the yield of full-length product
Saves time, because the enzyme blend requires less buffer optimization than single polymerase amplification systems and is less dependent on magnesium concentration than single polymerase PCR systems
Enhances results with difficult experimental systems, because the enzyme blend outperforms Taq DNA Polymerase by more accurately amplifying critical sequences (GC-rich sequences or repeat-rich sequences)
Ideal for multiplex PCR, because the enzyme blend produces more homogeneous fragment patterns and greater yields than Taq DNA Polymerase alone

Roche Contact Information

Roche

Roche Diagnostics Corporation
P.O. Box 50414
9115 Hague Road
Indianapolis, IN 46250-0414 USA

Customer Service: 800-262-1640

Tech Support: 800-262-4911

Fax Number: 800-428-2883

Web Site: http://www.roche-applied-science.com

Expand High Fidelity PCR System